{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,5,8]],"date-time":"2026-05-08T22:08:35Z","timestamp":1778278115695,"version":"3.51.4"},"reference-count":267,"publisher":"MDPI AG","issue":"12","license":[{"start":{"date-parts":[[2020,12,6]],"date-time":"2020-12-06T00:00:00Z","timestamp":1607212800000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"DOI":"10.13039\/501100006742","name":"Cooperativa de Ensino Superior Polit\u00e9cnico e Universit\u00e1rio","doi-asserted-by":"publisher","award":["IMPLDEBRIS-PL-3RL-IINFACTS-2019"],"award-info":[{"award-number":["IMPLDEBRIS-PL-3RL-IINFACTS-2019"]}],"id":[{"id":"10.13039\/501100006742","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100006742","name":"Cooperativa de Ensino Superior Polit\u00e9cnico e Universit\u00e1rio","doi-asserted-by":"publisher","award":["BD\/CBAS\/CESPU\/01\/2020"],"award-info":[{"award-number":["BD\/CBAS\/CESPU\/01\/2020"]}],"id":[{"id":"10.13039\/501100006742","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100001871","name":"Funda\u00e7\u00e3o para a Ci\u00eancia e a Tecnologia","doi-asserted-by":"publisher","award":["SFRH\/BD\/116167\/2016"],"award-info":[{"award-number":["SFRH\/BD\/116167\/2016"]}],"id":[{"id":"10.13039\/501100001871","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Pharmaceutics"],"abstract":"<jats:p>Most cancer biologists still rely on conventional two-dimensional (2D) monolayer culture techniques to test in vitro anti-tumor drugs prior to in vivo testing. However, the vast majority of promising preclinical drugs have no or weak efficacy in real patients with tumors, thereby delaying the discovery of successful therapeutics. This is because 2D culture lacks cell\u2013cell contacts and natural tumor microenvironment, important in tumor signaling and drug response, thereby resulting in a reduced malignant phenotype compared to the real tumor. In this sense, three-dimensional (3D) cultures of cancer cells that better recapitulate in vivo cell environments emerged as scientifically accurate and low cost cancer models for preclinical screening and testing of new drug candidates before moving to expensive and time-consuming animal models. Here, we provide a comprehensive overview of 3D tumor systems and highlight the strategies for spheroid construction and evaluation tools of targeted therapies, focusing on their applicability in cancer research. Examples of the applicability of 3D culture for the evaluation of the therapeutic efficacy of nanomedicines are discussed.<\/jats:p>","DOI":"10.3390\/pharmaceutics12121186","type":"journal-article","created":{"date-parts":[[2020,12,10]],"date-time":"2020-12-10T20:18:22Z","timestamp":1607631502000},"page":"1186","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":385,"title":["Three-Dimensional Spheroids as In Vitro Preclinical Models for Cancer Research"],"prefix":"10.3390","volume":"12","author":[{"ORCID":"https:\/\/orcid.org\/0000-0002-8804-6104","authenticated-orcid":false,"given":"B\u00e1rbara","family":"Pinto","sequence":"first","affiliation":[{"name":"Cooperativa de Ensino Superior Polit\u00e9cnico e Universit\u00e1rio (CESPU), Instituto de Investiga\u00e7\u00e3o e Forma\u00e7\u00e3o Avan\u00e7ada em Ci\u00eancias e Tecnologias da Sa\u00fade (IINFACTS), Instituto Universit\u00e1rio de Ci\u00eancias da Sa\u00fade (IUCS), 4585-116 Gandra PRD, Portugal"}]},{"given":"Ana C.","family":"Henriques","sequence":"additional","affiliation":[{"name":"Cooperativa de Ensino Superior Polit\u00e9cnico e Universit\u00e1rio (CESPU), Instituto de Investiga\u00e7\u00e3o e Forma\u00e7\u00e3o Avan\u00e7ada em Ci\u00eancias e Tecnologias da Sa\u00fade (IINFACTS), Instituto Universit\u00e1rio de Ci\u00eancias da Sa\u00fade (IUCS), 4585-116 Gandra PRD, Portugal"},{"name":"Instituto Nacional de Engenharia Biom\u00e9dica (INEB), Universidade do Porto, 4099-002 Porto, Portugal"}]},{"given":"Patr\u00edcia M. A.","family":"Silva","sequence":"additional","affiliation":[{"name":"Cooperativa de Ensino Superior Polit\u00e9cnico e Universit\u00e1rio (CESPU), Instituto de Investiga\u00e7\u00e3o e Forma\u00e7\u00e3o Avan\u00e7ada em Ci\u00eancias e Tecnologias da Sa\u00fade (IINFACTS), Instituto Universit\u00e1rio de Ci\u00eancias da Sa\u00fade (IUCS), 4585-116 Gandra PRD, Portugal"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-4006-5779","authenticated-orcid":false,"given":"Hassan","family":"Bousbaa","sequence":"additional","affiliation":[{"name":"Cooperativa de Ensino Superior Polit\u00e9cnico e Universit\u00e1rio (CESPU), Instituto de Investiga\u00e7\u00e3o e Forma\u00e7\u00e3o Avan\u00e7ada em Ci\u00eancias e Tecnologias da Sa\u00fade (IINFACTS), Instituto Universit\u00e1rio de Ci\u00eancias da Sa\u00fade (IUCS), 4585-116 Gandra PRD, Portugal"}]}],"member":"1968","published-online":{"date-parts":[[2020,12,6]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"253","DOI":"10.1038\/nrd3144","article-title":"Anticancer drug development: The grand challenges","volume":"9","author":"Hait","year":"2010","journal-title":"Nat. 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